Hematopoietic origins of fibroblasts: II. In vitro studies of fibroblasts, CFU-F, and fibrocytes
Autor: | Masahiro Masuya, Hitoshi Minamiguchi, Christopher J. Drake, Paul A. Fleming, Makio Ogawa, Yasuhiro Ebihara, Richard P. Visconti, Amanda C. LaRue |
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Rok vydání: | 2006 |
Předmět: |
Male
Cancer Research Green Fluorescent Proteins Population Vimentin In Vitro Techniques Biology Collagen Type I Colony-Forming Units Assay Mice Genetics medicine Animals Transplantation Homologous RNA Messenger education Fibroblast Discoidin Domain Receptors Molecular Biology Cells Cultured education.field_of_study Reverse Transcriptase Polymerase Chain Reaction Hematopoietic Stem Cell Transplantation Receptor Protein-Tyrosine Kinases Hematopoietic stem cell Cell Biology Hematology Fibroblasts Flow Cytometry Hematopoietic Stem Cells Molecular biology Clone Cells Fibronectins Collagen Type I alpha 1 Chain Mice Inbred C57BL Fibronectin Transplantation Haematopoiesis medicine.anatomical_structure Gene Expression Regulation Receptors Mitogen biology.protein Female Bone marrow |
Zdroj: | Experimental Hematology. 34:219-229 |
ISSN: | 0301-472X |
Popis: | Objective Using transplantation of a clonal population of cells derived from a single hematopoietic stem cell (HSC) of transgenic enhanced green fluorescent protein (EGFP) mice, we have documented the hematopoietic origin of myofibroblasts, such as kidney mesangial cells and brain microglial cells. Because myofibroblasts are thought to be an activated form of fibroblasts, we tested the hypothesis that fibroblasts are derived from HSCs. Materials and Methods Clones of cells derived from single cells of EGFP Ly-5.2 C57Bl/6 mice were transplanted into lethally irradiated Ly-5.1 mice. Using bone marrow and peripheral blood cells from mice showing high-level multilineage hematopoietic reconstitution, we induced growth of fibroblasts in vitro. Results Culture of EGFP + bone marrow cells from clonally engrafted mice revealed adherent cells with morphology typical of fibroblasts. Flow cytometric analysis revealed that the majority of these cells are CD45 − and express collagen-I and the collagen receptor, discoidin domain receptor 2 (DDR2). Reverse transcriptase polymerase chain reaction analysis of cultured cells demonstrated expression of procollagen 1-α1, DDR2, fibronectin, and vimentin mRNA. Fibroblast colonies consisting of EGFP + cells were observed in cultures of bone marrow cells from clonally engrafted mice, indicating an HSC origin of fibroblast colony-forming units. Culture of peripheral blood nucleated cells from clonally engrafted mice revealed EGFP + cells expressing collagen-I and DDR2, indicating that fibrocytes are also derived from HSCs. Conclusion We conclude that a population of fibroblasts and their precursors are derived from HSCs. |
Databáze: | OpenAIRE |
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